Drajad AS Penerapan teknologi inseminasi buatan, embrio transfer dan in vitro fertilisasi pada rusa Indonesia. Laporan Riset Unggulan Terpadu

(1)

DAFTAR PUSTAKA

Anderson LL. 2000. Pigs. In: Hafez ESE, Hafez B, editor. Reproduction in farm Animals.7

^th

Ed. USA: Williams & Wilkins.

Aritonang A. 1993. Babi, Perencanaan dan Pengelolaan Usaha. Bandung:

Penebar Swadaya.

Ax RL, Dally M, Didion BA, Lenz RW, Love CC, Varner DD, Hafez B, Bellin ME. 2000a. Semen Evaluation. In: Hafez ESE, Hafez B, editor.

Reproduction in farm Animals.7

^th

Ed. USA: Williams & Wilkins.

Ax RL, Dally M, Didion BA, Lenz RW, Love CC, Varner DD, Hafez B, Bellin ME. 2000b. Artificial Insemination. In: Hafez ESE, Hafez B, editor.

Reproduction in farm Animals.7

^th

Ed. USA: Williams & Wilkins.

Bassol J, Kádár E, Briz MD, Pinart E, Sancho S, Garcia-Gil N, Badia E, Pruneda A, Coll MG, Bussalleu E, Yeste M, Bonet S. 2005. In vitro culture of boar epididymal epithelial cells. Theriogenology 63: 363-369.

Bearden HJ, Fuquay JW. 1997. Applied animal reproduction. 4

^th

Ed. New Jersey:

Prentice Hall, Upper Saddle:Pp 133-177.

Beaulieu M, Dube C, Reyes-Moreno C, Guillemette C, Bailey J.L. 2005.

Differential effects of BTS and Androhep on boar semen [abstrack]. Di dalam : Gadella B.M & Colenbrander B, editor. Proceedings of the V International Conference on Boar Semen Preservation; Doorwerth, The Netherlands, 24-27 August 2003. Theriogenology 63: 422-430.

Blakely J, Bade DH. 1985. Ilmu Peternakan Ed.4. Srigandono B, penerjemah;

Yogyakarta: Gadjah Mada University Press. Terjemahan dari Animal Husbandry 4

^th

Ed.

Bonet S, Briz M, Fradera A. 1993. Ultrastructural abnormalities of boar spermatozoa. Theriogenology 40: 383-396

Bruce A, Dennis B, Julian L, Martin R, Keith R. 1994, Molecular Biology of The Cell, 3

^rd

Ed., New York: Garland Publishing, Inc.

Chun-Xia Z, Zeng-Ming Y. 2000. Evaluation on sperm quality of freshly ejaculated boar semen during in vitro storage under different temperatures.

Theriogenology 53(7): 1477-1488.

Coffey RD, Parker GR, Laurent KM. 2007. Manipulation of the Estrous Cycle in Swine.

www.uky.edu/Ag/AnimalSciences/pubs/asc152.pdf

(1 Agustus 2007)

Colenbrander B. Fazeli AR. Van Buiten A. Parlevliet J. Gadella BM. 1992.

Assesment of sperm cell membran integrity in the horse. Act Vet Scand.

Supl. 88 : 49-58.

De Leeuw FE, Colenbrander B, Verkleij AJ. 1990. The role membrane damage

plays in cold shock and freezing injury. Reprod. Domest. Anim. 1: 95-104.

(2)

Drajad AS. 1994. Penerapan teknologi inseminasi buatan, embrio transfer dan in vitro fertilisasi pada rusa Indonesia. Laporan Riset Unggulan Terpadu V bidang Teknologi Perlindungan Lingkungan. Hlm:92-111.

Dube C, Beaulieu M, Reyes-Moreno C, Guillemette C, Bailey J.L. 2004. Boar sperm storage capacity of BTS and Androhep Plus: viability, motility, capacitation, and tyrosine phosphorylation. Theriogenology 62: 874-886.

Evans G, Maxwel WMC. 1987. Salamon’s Artificial Insemination of Sheep and Goat. Sydney: Butterworths.

Everett RW, Bean B. 1982. Environmental influence on semen output. J Dairy Sci. 65:1303-1310

Fisrt NL. 1970. Collection, Evaluation and Insemination of Boar Semen. Dep.

Meat and Anim Sci. Winconsin University, Madison.

Frandson. 1965. Anatomy & Physiology of Farm Animals. Philadelphia : Lea &

Febiger.

Gadea J. 2003. Semen extenders used in the artificial insemination of swine.

Spanish Journal of Agricultural Research 1 (2): 17-27.

Garcia-Casado P, Marigorta P, Diaz C, Saiz-Cidoncha F. 2005. Use of the dehydrated egg yolk for boar semen freezing [abstrack]. Di dalam : Gadella B.M & Colenbrander B, editor. Proceedings of the V International Conference on Boar Semen Preservation; Doorwerth, The Netherlands, 24- 27 August 2003. Theriogenology 63: 422-430.

Garcia MA, Graham EF. 1989. Development of buffer system analysis of bovine spermatozoa before freezing. II. Effects of sugars and sugar alcohols on post thaw motility. Theriogenology 31: 1029-1037.

Garner DL, Hafez ESE. 2000. Spermatozoa and Seminal Plasma. In: Hafez ESE, Hafez B, editor. Reproduction in farm Animals.7

^th

Ed. USA: Williams &

Wilkins.

Girisonta. 1981. Pedoman Lengkap Beternak Babi. Yogyakarta: Kanisius.

Gunarso W. 1989. Mikroteknik. Bogor: Pusat Antar Universitas Ilmu Hayat Institut Pertanian Bogor.

Hafez B, Hafez ESE. 2000. Reproductive Behavior. In: Hafez ESE, Hafez B, editor. Reproduction in farm Animals.7

^th

Ed. USA: Williams & Wilkins.

Hirai M, Boersma A, Hoeflich A, Wolf E, Foll J, Aumuller TR, Braun J. 2001.

Objectively measured sperm motility and sperm head morphometry in boars (Sus scrofa): relation to fertility and seminal plasma growth factors. J Androl 22: 104-110.

Hirotada T, Emi O, Abdul GM, Sharoare H, Ummay S. 2006. Effect of fructose on motility, acrosome reaction and invitro fertilization capability of boar spermatozoa. Reproductive Medicine and Biology 5(4): 255-261

Huo Li-Jun, Ma Xing-Hog, Yang Zeng-Ming. 2002. Assesment of sperm

viability, mitochondrial activity, capacitation and acrosome intacness in

extender boar during long-term storage. Theriogenology 58(7): 1349-1360.

(3)

Jainudeen MR, Hafez ESE. 2000. Gestation, Prenatal Physiology, and Parturition.

In: Hafez ESE, Hafez B, editor. Reproduction in farm Animals.7

^th

Ed. USA:

Williams & Wilkins.

Johnson L.A, Aalbers J.G, Willems C.M.T, Rademaker J.H.M, Rexroad C.E.

1982. Use of boar spermatozoa for artificial insemination : bagian III Fecundity of boar spermatozoa stored in Beltsville liquid and Kiev extenders for three days at 18°C. J Anim Sci 54(1): 132-136

Johnson LA, Weitze KF, Fiser P, Maxwell WMC. 2000. Storage of boar semen. J Anim Sci 62: 143-172.

Kadirvel G, Nasker S, Das A, Hasin D. 2005. Effect of different extenders on preservation of boar semen at 17°C [abstrack]. Di dalam : Gadella B.M &

Colenbrander B, editor. Proceedings of the V International Conference on Boar Semen Preservation; Doorwerth, The Netherlands, 24-27 August 2003.

Theriogenology 63: 685-692.

Kevin R. 2000. Fresh Boar Semen. In: Swine News. College of Agriculture &

Life Sciences. 23: 11

Kommisrud E, Paulenz H, Sehested E, Grevle IS. 2002. Influence of boar and semen parameters on motility and acrosome integrity in liquid boar semen stored for five days. Acta Vet Scand 43(1): 49-55

Li-Jun H, Xing-Hong M, Zeng-Ming Y. 2002. Assesment of sperm viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-term storage. Theriogenology 58: 1349-1360.

Maxwell WMC, Salamon S. 1993. Liquid storage of ram semen. Reprod Fertil Dev. 5:613-638.

Molinia FC, Evans G, Maxwell WMC. 1994. In vitro evaluation of zwitterion buffer in diluent for freezing ram spermatozoa. Reprod. Nutr. Dev. 34: 491- 500.

Morel DMCG. 1999. Equine Artificial Insemination. CABI Publishing, Wallingford, Oxon, UK.

Parks JE, Graham JK. 1992. Effects of cryoprotectant procedures on sperm membran. Theriogenology 38: 385-392.

Paulenz H, Kommisrud E, Hofmo PO. 2000. Effect of long-term storage at different tempertures on the quality of liquid boar semen. Reprod Dom Anim 35: 83-85.

Ponglowhapan S, Gustavsson BE, Forsbeg CL. 2004. Influence of glucose and fructose in the extender during long-term storage of chilled canine semen.

Theriogenology 62: 1498-1517.

Pursel VG, Johnson LA, Schulman LL. 1973. Effect of dilution, seminal plasma

and incubating period on cold shock susceptibility of boar spermatozoa. J

Anim Sci 37: 528-531

(4)

Rigau T, Piedrafita J, Reverter J, Canal M, Rodriguez-Gil JE. 1996. The rate of L- lactate production: a feasible parameter for the fresh diluted boar semen quality analysis. Anim Reprod Sci. 43: 161-172

Robert VK. 2006. Semen Processing, Extending & Storage for Artificial Insemination in Swine. Dep. of Animal Science University of Illinois.

Sihombing DTH. 2006. Ilmu Ternak Babi. Yogyakarta: Gadjah Mada University Press.

Sorenson AM. 1979. A Laboratory Manual for Animal Reproduction 4

^th

Ed.

Texas: Animal Science Departemen A & M University.

Shukla SN, Sigh BB, Tomar NS, Misra BS. 1992. Factor effecting spermatozoa motility in preserved semen. J. Indian Vet. 69: 856-857.

Singleton WL. 2001. State of the art in artificial insemination in the United States.

Theriogenology 56: 1305-1310.

Supriatna I, Pasaribu FH. 1992. In vitro fertilisasi, transfer embrio dan pembekuan embrio. Bogor: Pusat Antar Universitas Bioteknologi, Institut Pertanian Bogor.

Steinbach J, Foote RH. 1967. Osmotic pressure and pH effects on survival of frozen or liquid spermatozoa. J. Dairy Sci. 50:205.

Stell RGD, Torrie JH. 1993. Principles and Procedures of Statistics. 2

^th

Ed.

London: International Student Edition.

Sterle J, Safranski T. 1997. Artificial Insemination in Swine : Breeding the Female. Dept. of Anim. Sci. University of Missouri

Toelihere MR. 1993. Inseminasi Buatan pada Ternak. Bandung: Angkasa.

Toelihere MR. 1981. Fisiologi Reproduksi pada Ternak. Bandung: Angkasa.

Trejo AG, Anaya MJ, Hermandez GM. 1996. Effect of the egg yolk concentration and the cooling rates on the sperm motility and acrosomal integrity of frozen caprine semen. VI International Conference on Goat. Vol 2. International Academic Publishers.

Vyt P, Maes D, Dejonckheere E, Castryck F, Van Soom A. 2004. Comparative study on five different commercial extenders for boar semen. Reprod Dom Anim 39(1): 8

Vyt P, Maes D, Dejonckheere E, Castryck F, Van Soom A. 2005. Comparison of porcine semen quality during one week of preservation using 5 commercial extenders [abstrack]. Di dalam : Gadella B.M & Colenbrander B, editor.

Proceedings of the V International Conference on Boar Semen Preservation;

Doorwerth, The Netherlands, 24-27 August 2003. Theriogenology 63: 685- 692.

Waberski D, Weitze KF, Rath D, Salman HP. 1989. Effect of bovine serum

albumin and zwitterionic buffers on stored liquid boar semen. Zuchthygiene

24: 128-133.

(5)

Waberski D, Wetze KF, Lietmann C, Lubbert W. 1994. The initial fertilizing capacity of longterm-stored liquid boar semen following pre-and postovulatory insemination. Theriogenology 41: 1367-1377.

Watson PF. 1990. Artificial insemination and the preservation of semen. Di dalam : Lamming G, editor. Marshall’s Physiology of Reproduction 4

^th

Ed., Churchill livingstone, Edinburgh, 2: 747-869.

Watson PF. 1995. Recent development and concept in the cryopreservation of spermatozoa and assement of their post-thawing function. Reprod Fertil Dev 7: 871-891.

Watson PF. 1996. Cooling of spermatozoa and fertilizing capacity. Reprod.

Domest. Anim. 31: 135-140.

White IG. 1993. Lipid and calcium uptake of sperm in relation to cold shock and preservation : A review. Reprod Fertil Dev 5: 639-658.

Yildiz C, Kaya A, Aksoy, Tekeli. 2000. Influence of sugar supplementation of extender on motility, viability and acrosomal integrity of dog spermatozoa during freezing. Theriogenoloy 54: 507-650.

Zhou J.B, Yuek K.Z, Luo M.J, Chang Z.L, Liang H, Wang Z.Y, Tan J.H. 2004.

Effect of extender and temperatures on sperm viability and fertility capacity

of harbin white boar semen during long-term liquid storage. Asian-

Australas.J.Anim.Sci 17(11): 1501-1508.

(6)

Lampiran 1 Persentase motilitas spermatozoa dari pejantan 1

Ruang Terbuka Kotak Styrofoam Lemari Es

Pengamatan

jam ke- TP BTS MBTS MZOC TP BTS MBTS MZOC TP BTS MBTS MZOC

0

63.33

63.33 6

36.67

56.67

60.00

58.33

61.67

60.00

61.67

58.33

61.67

61.67 12

25.83

51.67

49.17

58.33

53.33

60.00

56.67

60.00

53.33

60.00

60.00 18

15.00

46.67

41.67

56.67

41.67

56.67

48.33

60.00

41.67

59.17

55.00

58.33 24

5.00

33.33

15.00

43.33

30.00

53.33

40.00

60.00

30.00

58.33

50.00

56.67 30

-

16.67

5.00

40.00

15.00

46.67

31.67

55.00

15.00

54.17

40.00

51.67 36

-

5.00

-

32.33

5.00

30.00

20.00

40.00

5.00

30.00

25.00

40.00 42

-

1.67

-

21.20

-

20.00

10.00

30.00

-

20.00

10.00

30.00 rataan

18.23

34.38

28.85

46.90

33.33

48.96

41.25

53.75

33.33

50.83

45.63

52.71

Ket : TP (tanpa pengencer); BTS (beltsville thawing solution); MBTS (modifikasi BTS);

MZoc (modifikasi Zorlesco)

Lampiran 2 Persentase motilitas spermatozoa dari pejantan 2

Pengamatan

0

68.33

68.33 6

56.67

61.67

60.00

65.00

62.50

65.00

62.50

65.83

59.17

65.00

58.33

65.83 12

50.00

59.17

55.00

62.50

56.67

61.67

56.67

63.33

50.00

61.67

48.33

63.33 18

43.33

56.67

50.00

60.00

43.33

55.83

53.33

61.67

40.00

57.50

50.83

60.00 24

30.00

46.67

33.33

53.33

30.00

50.00

60.00

30.00

53.33

56.67 30

5.00

31.67

23.33

45.00

20.00

41.67

40.83

56.67

21.67

47.50

35.00

53.33 36

-

20.00

10.00

35.00

5.00

25.00

20.00

40.00

5.00

25.50

28.67

40.00 42

-

10.00

5.00

30.00

-

15.00

10.00

30.00

-

15.00

15.70

30.00 rataan

42.22

44.27

38.13

52.40

35.73

47.81

45.21

55.73

34.27

49.23

44.82

54.69

Lampiran 3 Persentase motilitas spermatozoa dari pejantan 3

Pengamatan

0

65.00

65.00 6

53.33

58.33

55.00

63.33

60.83

64.17

62.50

63.33

59.17

63.33

61.67

63.33 12

46.67

55.00

50.83

62.50

56.67

63.33

60.00

61.67

53.33

61.67

58.33

61.67 18

40.00

51.67

46.67

61.67

50.00

60.00

55.00

60.83

50.00

58.33

53.33

61.67 24

23.33

43.33

40.00

56.67

43.33

56.67

50.00

60.00

46.67

55.00

48.33

61.67 30

-

26.67

16.67

53.33

21.67

51.67

25.00

57.50

31.67

50.83

32.50

57.50 36

-

20.00

10.00

40.00

5.00

30.00

20.00

40.00

5.00

30.00

27.67

40.00 42

-

10.00

5.00

30.00

-

20.00

15.00

30.00

-

20.00

15.67

30.00 rataan

28.54

41.25

36.15

54.06

37.81

51.35

44.06

54.79

38.85

50.52

45.31

55.10

(7)

Lampiran 4 Persentase spermatozoa hidup dari pejantan 1

Pengamatan Ruang Terbuka Kotak Styrofoam Lemari Es

0

88.04

88.04 6

59.97

76.72

70.61

77.25

79.94

86.45

84.83

83.47

78.97

86.38

86.61

81.91 12

45.31

67.90

60.72

70.72

71.85

84.85

81.62

78.91

69.90

84.72

85.19

75.78 18

30.65

59.08

50.83

64.20

54.97

74.97

70.16

74.97

57.51

80.80

76.72

71.28 24

12.22

41.56

19.35

55.67

38.10

65.08

58.70

71.04

45.12

76.89

68.26

66.79 30

-

19.91

8.82

51.99

19.05

56.19

46.98

66.47

22.56

73.16

56.31

61.50 36

-

8.50

- - 10.00

45.00

25.00

50.00

10.00

40.00

28.65

45.00 42

-

5.00

- - -

25.00

15.00

35.00

-

25.00

15.00

35.00 rataan

29.52

45.84

37.30

50.98

45.24

65.70

58.79

68.49

46.51

69.37

63.10

65.66

Lampiran 5 Persentase spermatozoa hidup dari pejantan 2

0

90.48

90.48 6

73.95

80.77

78.96

79.62

79.89

85.05

82.76

85.24

77.95

85.83

84.17

86.04 12

66.20

75.74

70.20

72.63

69.29

79.63

75.03

80.00

65.42

81.17

77.86

81.60 18

58.46

70.71

61.44

65.64

54.34

71.63

68.56

76.71

52.75

76.21

68.58

75.72 24

37.42

50.33

44.82

61.47

39.39

63.64

62.09

73.41

40.08

71.25

59.29

69.85 30

12.22

40.12

30.97

54.44

27.61

52.51

51.07

69.50

30.75

60.54

40.26

64.19 36

-

27.67

15.00

47.67

10.00

30.00

22.50

48.67

10.00

33.67

30.00

45.50 42

-

15.00

8.50

40.00

-

22.50

12.22

35.50

-

20.00

20.50

35.00 rataan

42.34

56.35

50.04

63.99

46.37

61.93

58.09

69.94

45.93

64.89

58.89

68.55

Lampiran 6 Persentase spermatozoa hidup dari pejantan 3

0

84.76

84.76 6

64.10

68.54

68.76

78.77

74.98

79.12

76.38

81.07

77.54

78.74

77.10

81.43 12

55.74

65.11

61.99

76.47

65.20

73.47

68.00

77.38

70.31

72.73

69.44

78.09 18

47.38

61.68

55.23

74.17

59.85

68.39

63.43

75.49

65.56

66.72

65.10

76.62 24

26.67

51.62

47.92

62.05

54.50

63.30

58.86

73.61

60.81

60.71

60.75

75.14 30

-

30.68

17.36

57.19

27.25

59.91

29.43

69.67

40.04

54.78

39.96

68.02 36

-

25.00

15.00

42.50

10.00

35.00

27.67

47.67

12.22

43.50

30.00

45.67 42

-

12.22

8.50

32.00

-

27.50

18.67

38.50

-

27.67

18.67

33.67 rataan

34.83

49.95

44.94

63.49

47.07

61.43

53.40

68.52

51.41

61.20

55.72

67.93

(8)

Lampiran 7 Pengenceran semen Dari data yang diperoleh dilapangan : Volume semen : 214.44 mL Motilitas progresif : 65.56 %

Konsentrasi : 191.65 x 10

⁶

sel/mL Dapat dihitung :

volume x konsentrasi x motilitas

Volume total = x volume IB

konsentrasi IB

214.44 x (191.65 x 10

⁶

) x 65.56%

= x 80 mL

2000 x 10

⁶

(41097.43 x10

⁶

) x 65.56%

= x 80 mL

2000 x 10

⁶

26943.47 x 10

⁶

= x 80 mL = 1077.74 mL

2000 x 10

⁶

Volume pengencer = Volume total – volume semen

= 1077.74 – 214.44

= 863.3 mL

Dari perhitungan diatas dapat dilihat :

1. Total spermatozoa per ejakulat = 41097.43 x10

⁶

sel 2. Total spermatozoa motil per ejakulat = 26943.47 x 10

⁶

sel 3. Konsentrasi per ml setelah diencerkan

26943.47 x 10

⁶

sel

= x 80 mL

863.3 mL

= 2496.80 x 10

⁶

sel/80 mL

Jika dalam sembilan jam penyimpanan terjadi penurunan %M, maka dosis atau volume IB harus ditingkatkan untuk mencapai dosis standar IB yakni motilitas 65% dan konsentrasi 2 x 10

⁹

sel/80mL, dengan perhitungan :

= ( Motilitas IB / Motilitas semen ) x Volume IB

= ( 65% / 40%) x 80 mL

= 130 mL

Jadi, dengan motilitas 40% dan konsentrasi 2 x 10

⁹

, volume IB yang digunakan

adalah 130 mL.

(9)

Lampiran 8 Rangkaian kegiatan IB pada babi 1. Prosesing Semen Cair

Persiapan bahan pengencer Persiapan dummy sow

Persiapan pejantan Persiapan alat penampungan semen

Penampungan semen Evaluasi dan Pengenceran

Penyimpanan Paking

Dilanjutkan dengan pelaksanaan inseminasi,

seperti yang diperlihatkan pada halaman dibawah ini.

(10)

2. Pelaksanaan Inseminasi Buatan a. Deteksi berahi

Vulva Merah Vulva hangat (35-38 °C)

Uji tekan punggung Vulva berlendir

b. Persiapan alat inseminasi

Kateter (50-55 cm) Kemasan semen (80 ml) dengan ujung berbentuk spiral

Ujung

spiral

(11)